Lipid classes and their content of n-3 highly unsaturated fatty acids (HUFA) in Artemia franciscana after hatching, HUFA-enrichment and subsequent starvation

The distribution of n-3 highly unsaturated fatty acids (HUFA) over the major neutral and polar lipid classes was determined for two predominant types of live food used in the larviculture of marine fish and shrimp, i.e. freshly hatched and HUFA-enriched Artemia, and compared with data reported in the literature for wild copepods, representing the natural diet of these larvae. Lipid class composition and their content of n-3 HUFA, particularly eicosapentaenoic acid (EPA, 20:5n-3) and docosahexaenoic acid (DHA, 22:6n-3), were assessed in freshly hatched, HUFA-enriched and subsequently starved Artemia franciscana. The n-3 HUFA enrichment was based on feeding Artemia a lipid emulsion in which either fatty acid ethyl esters (EE, diluted with olive oil) or triacylglycerol (TAG) provided a level of 30% n-3 HUFA. Enrichment of Artemia with either type of the lipid emulsions resulted in an increase of total lipid content from 20.0 to 28.2–28.7% of dry matter mainly due to the accumulation of neutral lipid, primarily TAG (from 82 to 158 mg g⁻¹ dry wt in freshly hatched and 24-h enriched Artemia). Enriched brine shrimp utilized up to 27–30% of their TAG content during 72 h of starvation at 12 °C. The absolute tissue concentrations of polar lipids remained constant at 71 to 79 mg g⁻¹ dry wt throughout the enrichment and subsequent starvation. The level of n-3 HUFA increased drastically during enrichment from 6.3% of total fatty acids (8.2 mg g⁻¹ dry wt) in freshly hatched nauplii to between 20.4 and 21.8% (40.4 to 43.2 mg g⁻¹ dry wt) in 24-h enriched Artemia and was not significantly affected by the source of n-3 HUFA. During starvation, 18:0, 20:4n-6 and 20:5n-3 were retained, whereas 18:4n-3, 22:5n-3 and 22:6n-3 were specifically catabolized. The major polar lipids, phosphatidylethanolamine (PE) and phosphatidylcholine (PC), of freshly hatched Artemia showed very low levels of DHA (<0.1% of total fatty acids) and carried about 45% of the total EPA present. Enrichment with either of the emulsions resulted in an increase of the neutral lipid fraction which concentrated >64% of the EPA and >91% of the total DHA present. This is in sharp contrast with the high levels of n-3 HUFA, in particular DHA, in the polar lipid fraction reported for wild copepods. The contrasting distribution of DHA in the neutral and polar lipid fractions of enriched brine shrimp compared to the natural diet may influence the efficacy of this essential fatty acid for marine fish larvae in aquaculture systems. Received: 10 June 1997 / Accepted: 8 August 1997     

Effects of monospecific and mixed-algae diets on survival, development and fatty acid composition of penaeid prawn (Penaeus spp.) larvae

Four species of microalgae (Chaetoceros muelleri, Tetraselmis suecica, Tahitian Isochrysis sp. (T-iso) and Dunaliella tertiolecta) with distinctly different fatty acid profiles were grown in continuous culture and fed to prawn larvae (Penaeus japonicus, P. semisulcatus and P. monodon) as monospecific diets. The best two diets (C. muelleri and T. suecica) were also fed as a mixed diet. Experiments were run until the larvae fed the control diet of C. muelleri metamorphosed to Mysis 1. The survival and development (i.e. performance) of the larvae were affected by algal diet, and the diets were ranked in the order of decreasing nutritional value: C. muelleri ≥ T. suecica > T-iso > D. tertiolecta. Larvae fed a mixed diet of C. muelleri and T. suecica (2:3 by dry weight) performed as well or better than those fed C. muelleri, and the performance of both these groups of larvae was better than those fed T. suecica. The lipid and carbohydrate compositions of the algae had little or no effect on the lipid and carbohydrate compositions of the larvae or their performance. However, the larvae that performed best (i.e. those fed C. muelleri) had significantly more lipid and carbohydrate than those that performed worst (i.e. those fed D. tertiolecta). Larvae fed C. muelleri or the mixed-algae diet had higher proportions of the essential fatty acids eicosapentaenoic acid [EPA, 20:5(n-3)] and arachidonic acid [ARA, 20:4(n-6)] than the larvae fed on other diets. Furthermore, the larvae fed T. suecica, which showed intermediate performance between larvae fed C. muelleri and T-iso or D. tertiolecta, also had higher proportions of EPA and ARA. Both C. muelleri and T. suecica contained EPA and ARA, but T-iso and D. tertiolecta did not, except for trace amounts of EPA in T-iso. The fatty acid ARA appears to be much more important in the diet of larval prawns than has so far been considered. The level of the essential fatty acid docosahexaenoic acid [DHA, 22:6(n-3)] in the algal diet and the larvae was not related to the performance of the larvae; only C. muelleri and T-iso contained DHA. However, the nauplii contained large proportions of DHA, suggesting that these were sufficient to meet the larval requirements for DHA during their development to Mysis 1. Mixed-algae diets could improve the performance of larvae by providing a more comprehensive range of fatty acids. Received: 22 April 1998 / Accepted: 3 December 1998     

Effects of salinity and dietary DHA (22:6n - 3) content on lipid composition and performance of Penaeus kerathurus postlarvae

A two-way ANOVA experiment was designed to study the effects of salinity and dietary docosahexaenoic acid (DHA; 22:6n-3) on lipid composition and performance of postlarvae from the marine shrimp Penaeus kerathurus (Forskål, 1775). Shrimp were reared from 1- to 8-d-old postlarvae at 35 and 25‰S with Kelko-enriched Artemia sp. [20.0 μg (n-3)?HUFA mg^?1 dry weight; 9.1 μg DHA mg^?1 dry weight] and nonenriched Artemia sp. [14.2 μg (n-3)?HUFA mg^?1 dry weight; 0.3 μg DHA mg^?1 dry weight]. Dietary DHA content did not affect either total length or survival but influenced the nutritional status represented by condition indices (triacylglycerol/total polar lipid and triacylglycerol/free cholesterol) of 8-d-old postlarvae at the end of the experiment. Culture salinity affected final total length and condition indices but did not show any effect on survival in the different experimental treatments. The interaction of dietary DHA and culture salinity was not significant for total length and survival but was significant for both condition indices used. P. kerathurus 8-d-old postlarvae showed better growth, survival and nutritional condition when reared at 35‰S and when fed on Kelko-enriched Artemia sp., but the differences with postlarvae from other treatments were very poorly marked. The results demonstrate that 8-d-old postlarvae may have sufficiently developed osmoregulatory capabilities to resist 25‰S under good conditions, although (n-3) HUFA-enriched diets may also enhance osmotic stress resistance, general performance and disease resistance.     

Effects of algae-enriched rotifers on winter flounder (Pseudopleuronectes americanus) gene expression during metamorphosis

The aim of this study was to evaluate the effect of a dietary highly unsaturated fatty acid (HUFA) deficiency on winter flounder Pseudopleuronectes americanus metamorphosis by examining the growth and the expression of genes involved in some key metabolic processes: lipid digestion, oxidative stress, and growth. Three groups of fish were fed rotifers enriched with different blends of microalgae providing different HUFA profiles: (1) a diet comprising a mixture of three microalgae, Nannochloropsis oculata, Isochrysis galbana, and Pavlova lutheri (Cocktail diet), which contained a balanced combination of ecosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and arachidonic acid (AA); (2) the N. oculata diet (Nanno diet), with a low level of DHA; and (3) the I. galbana diet (Tiso diet), characterized by low levels of EPA and AA. The results indicate that the need for DHA increased from settlement and for EPA and AA from 15 days after settlement. The lower HUFA content in the Tiso and Nanno diets did not affect larval development or lipid reserve accumulation. The superoxide dismutase gene expression suggests a reduced oxidative stress in the Cocktail group, and overall results indicate that gh gene expression could be a valuable indicator of development at the molecular level in response to dietary HUFA quality during metamorphosis in winter flounder.     

An experimental study of the effect of diet on the fatty acid profiles of the European Cuttlefish (<Emphasis Type="Italic">Sepia officinalis</Emphasis>)

Fatty acid analysis is an alternative dietary investigation tool that complements the more traditional techniques of stomach content and faecal analysis that are often subject to a wide range of biases. In applying fatty acid analysis to ecosystem studies, it is important to have an understanding of the effect diet has on the fatty acid profile of the predator. A feeding experiment, using crustacean and fish as prey for the European cuttlefish Sepia officinalis, was conducted to evaluate the effect of prey fatty acids on the fatty acid profile of this marine predator. Cuttlefish were fed on a fish diet for the first 29 days, and then changed to a crustacean diet for a further 28 days. Another group of cuttlefish was fed on a crustacean diet for the first 29 days, and then changed to a fish diet for a further 28 days. An analysis of the cuttlefish digestive gland showed that the fatty acid profile reflected that of the prey, with cuttlefish on a crustacean diet being clearly distinguishable from the cuttlefish on a fish diet. Cuttlefish fed on a fish diet for 29 days prior to the switch in diet were comparatively higher in 16:0, AA, 20:1ω9, DPA6, DHA, 22:4ω6 and DPA3 than those fed on crustaceans. Cuttlefish fed on a crustacean diet for 29 days prior to the switch in diet were comparatively higher in 17:1ω8, 18:1ω9, 18:2ω6, 18:1ω7, EPA and 20:2ω6 than those fed on fish. Following a change in diet, the fatty acid profile of the cuttlefish digestive gland reflected that of the new diet within 14 days. The results confirm that the fatty acid profile of the cuttlefish digestive gland clearly reflects the profile of its recent diet. It also shows that the digestive gland may not be an organ that accumulates dietary lipids for long-term storage, but rather is an organ where lipids are rapidly being turned over and potentially excreted.     

Essential fatty acids influence metabolic rate and tolerance of hypoxia in Dover sole (Solea solea) larvae and juveniles

Dover sole (Solea solea, Linneaus 1758) were raised from first feeding on brine shrimp (Artemia sp.) with different contents and compositions of the essential fatty acids (EFA) arachidonic acid (ARA, 20:4n − 6); eicosapentaenoic acid (EPA, 20:5n − 3), and docosahexaenoic acid (DHA, 22:6n − 3), and their metabolic rate and tolerance to hypoxia measured prior to and following metamorphosis and settlement. Four dietary Artemia preparations were compared: (1) un-enriched; (2) enriched with a commercial EFA mixture (Easy DHA SELCO Emulsion); (3) enriched with a marine fish oil combination (VEVODAR and Incromega DHA) to provide a high ratio of ARA to DHA, and (4) enriched with these fish oils to provide a low ratio of ARA to DHA. Sole fed un-enriched Artemia were significantly less tolerant to hypoxia than the other dietary groups. Larvae from this group had significantly higher routine metabolic rate (RMR) in normoxia, and significantly higher O₂ partial pressure (PO₂) thresholds in progressive hypoxia for their regulation of RMR (P _crit) and for the onset of agitation, respiratory distress and loss of equilibrium. Metamorphosis was associated with an overall decline in RMR and increase in P _crit, but juveniles fed on un-enriched Artemia still exhibited higher P _crit and agitation thresholds than the other groups. Sole fed un-enriched Artemia had significantly lower contents of EFA in their tissues, both before and after settlement. Thus, enriching live feeds with EFA has significant effects on the respiratory physiology of sole early life stages and improves their in vivo tolerance to hypoxia. We found no evidence, however, for any effect of the ratio of ARA to DHA.     

Effect of cobalt-supplemented diets on bioaccumulation,digestive enzyme activities and growth of Cyprinus carpio

Experiments were conducted to determine the effects of excess dietary cobalt (Co) on bioaccumulation, digestive enzyme activities, and growth in freshwater Cyprinus carpio. Four isonitrogenous diets (average crude protein: 35%) were formulated to prepare a control diet (T₁) with no Co, and three Co-supplemented diets with 1 (T₂), 1.5 (T₃), or 2 (T₄)% Co. The results showed that C. carpio fed with T₃ diet showed maximal apparent protein digestibility, feed conversion, protein utilization, and growth. Protease and lipase activities were maximal in the fish group given T₃ diet. Accumulation of Co in different soft and hard tissues of fish did not correlate with levels of Co in the diet. The concentration of Co in the water increased with dietary level, being at a disturbing adverse effect level at 2% dietary Co. Evidence indicates that an additional supply of dietary Co up to a level of 1.5% is a viable option to augment growth of C. carpio, but at higher levels of Co it may be detrimental to fish health and the aquatic ecosystem.     

Reduced n-3 highly unsaturated fatty acids dietary content expected with global change reduces the metabolic capacity of the golden grey mullet

In this study, we hypothesised that a reduction in n-3 HUFA availability for higher consumers, as expected with global change, would negatively impact the physiological performances of fish. The aim was to experimentally evaluate the effect of n-3 HUFA dietary content on cardio-respiratory performances of the golden grey mullet (Liza aurata), a microalgae grazer of high ecological importance in European coastal areas. These performances were evaluated in terms of critical swimming speed U _crit, associated oxygen consumption MO₂, post-exercise oxygen consumption and calcium fluxes in cardiomyocytes. Two replicated groups of fish were fed on a rich (standard diet, SD diet: 1.2 % n-3 HUFA on dry matter basis, DMB) or a poor n-3 HUFA (low n-3 HUFA diet, LD diet: 0.2 % n-3 HUFA on DMB) diet during 5 months and were called SD and LD groups, respectively. The results showed that the LD diet reduced growth rate as well as the aerobic capacity of L. aurata at 20 °C, suggesting that fish may have to save energy by modifying the proportion of energy allocated to energy-demanding activities, such as digestion or feeding. In addition, this LD diet induced higher levels of haematocrit and plasma osmolality, indicating a stress response at the second and third levels in that group. However, the LD diet caused a massive increase in swimming efficiency. This should improve the capacity of L. aurata to migrate and to forage over a wide area. In turn, these could then compensate for the reduction in growth rate and aerobic metabolism.     

Bioaccumulation of cobalt,digestive enzyme activities,and growth of freshwater catfish,Heteropneustes fossilis (Bloch), fed cobalt-supplemented diets

Experiments were conducted to determine the effects of excess dietary cobalt (Co) on its bioaccumulation, on digestive enzyme activities, and on the growth of freshwater catfish Heteropneustes fossilis (Bloch). Four isonitrogenous diets (average crude protein: 32.7%) were formulated to prepare a control diet (T1) with no Co, and three Co-supplemented diets with 0.05 (T2), 0.1 (T3), and 1.0 (T4)% Co. The results showed that H. fossilis fed with T3 diet showed maximum apparent protein digestibility, feed conversion, protein utilization, and growth. Protease and lipase activities were also maximum in T3 diet. Accumulation of Co in different soft and hard tissues of the fish did not show any correlation with the level of Co in the diet. The concentration of Co in the water increased with its dietary level, being at an alarming level at 1.0% dietary Co. It is concluded that an additional supply of dietary Co up to a level of 0.1% is a viable option to augment growth of the catfish H. fossilis, but at higher levels of Co it may be detrimental to the fish and the aquatic ecosystem.     

Trophic modification of essential fatty acids by heterotrophic protists and its effects on the fatty acid composition of the copepod Acartia tonsa

To test whether heterotrophic protists modify precursors of long chain n−3 polyunsaturated fatty acids (LCn−3PUFAs) present in the algae they eat, two algae with different fatty acid contents (Rhodomonas salina and Dunaliella tertiolecta) were fed to the heterotrophic protists Oxyrrhis marina Dujardin and Gyrodinium dominans Hulbert. These experiments were conducted in August 2004. Both predators and prey were analyzed for fatty acid composition. To further test the effects of trophic upgrading, the calanoid copepod Acartia tonsa Dana was fed R. salina, D. tertiolecta, or O. marina that had been growing on D. tertiolecta (OM-DT) in March 2005. Our results show that trophic upgrading was species-specific. The presence of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the heterotrophic protists despite the lack of these fatty acids in the algal prey suggests that protists have the ability to elongate and desaturate 18:3 (n−3), a precursor of LCn−3PUFAs, to EPA and/or DHA. A lower content of these fatty acids was detected in protists that were fed good-quality algae. Feeding experiments with A. tonsa showed that copepods fed D. tertiolecta had a significantly lower content of EPA and DHA than those fed OM-DT. The concentration of EPA was low on both diets, while DHA content was highest in A. tonsa fed R. salina and OM-DT. These results suggest that O. marina was able to trophically upgrade the nutritional quality of the poor-quality alga, and efficiently supplied DHA to the next trophic level. The low amount of EPA in A. tonsa suggests EPA may be catabolized by the copepod.     

Effect of lipid emulsions on production and fatty acid composition of eggs of the scallop<Emphasis Type="Italic"> Argopecten purpuratus</Emphasis>

The impact of supplementing lipid emulsions rich in eicosapentaenoic acid (EmEPA), docosahexaenoic acid (EmDHA) or saturated fatty acids (EmCOCO) to a standard algal diet [3:1 mixture of Isochrysis galbana (T-iso) and Chaetoceros neogracile, St-diet] on Argopecten purpuratus broodstock was evaluated. Broodstock fecundity was compared as well as the egg quality in terms of lipid content, fatty acid composition and lipid class distribution. Fecundity was defined as the number of eggs released in the spawning process, since spawning was virtually complete. Results indicated that the total lipid content of the eggs of A. purpuratus was diet independent. A greater energy reserve was spent on a larger number of oocytes and not on bigger sized oocytes with a higher lipid content. The lipids supplied through the emulsions were at least partially allocated to the eggs, demonstrating that the fatty acid composition of the eggs could be manipulated, especially the neutral lipid fraction. Levels of EPA changed more rapidly than DHA levels, supporting the observation that they fulfilled an energetic and structural role, respectively. The St-diet supplemented with 50%EmCOCO resulted in a significantly higher fecundity compared to the algal diet supplemented with 25%EmEPA+25%EmDHA and the non-supplemented algal diet. It would seem that saturated fatty acids (SAFA) were more easily or preferentially incorporated in the female gonads of A. purpuratus. The relative content of SAFA and 18:2( n-6) in these eggs rose significantly. The relative content of the highly unsaturated fatty acids, EPA and DHA, on the other hand was substantially lower in the neutral lipid fraction, but hardly affected in the polar lipid fraction. It appeared that the maintenance of an adequate DHA/EPA ratio (approximately 1.2) was more important than the absolute levels of the two fatty acids, as long as a threshold value was reached.     

Membrane lipids of red muscle mitochondria from land-locked and sea-run Arctic char, Salvelinus alpinus

Although laboratory studies of the effects of temperature, salinity, and diet on biological membranes of fish indicate substantial alterations in phospholipid and fatty acid composition to maintain functional properties, there are few parallel studies of wild populations. We, therefore, examined the red muscle, mitochondrial phospholipids and phospholipid fatty acids in two fish populations differing in their environmental temperature, salinity, and diet. Sea-run and freshwater (land-locked) Arctic char (Salvelinus alpinus L.) were collected from Igloolik Island, Northwest Territories, Canada, in the summer of 1991. Several differences between the phospholipids of these fish, and those reported for red muscle mitochondria in other fish species, included a higher cardiolipin content and a higher proportion of short-chain monoenes, especially 16:1. In congruence with previous studies of changes in cardiolipin fatty acids in other species of cold-acclimated fish, the fatty acid content of cardiolipin of both Arctic char groups was more saturated and less polyunsaturated than in warm-acclimated fish. Other aspects of the lipid composition of these membranes were not consistent with laboratory studies of cold-acclimated fish. For example, the fatty acids comprising phosphatidylethanolamine and phosphatidylcholine were more saturated than would be predicted based on laboratory studies of cold-acclimated fish. Some of these differences may be attributable to differences in the proportions of dietary n3 and n6 fatty acids in freshwater and marine environments. A strategy common to both groups of Arctic char is the maintenance of a similar relationship between phospholipid fatty acid chain length and degree of unsaturation in both Arctic char populations in spite of differences in diet and thermal regimes. The observed differences in membrane composition between land-locked and sea-run fish presumably act to maintain mitochondrial function in these different environments. Received: 27 May 1997 / Accepted: 12 June 1997     

Growth and lipid composition of scallop juveniles, Placopecten magellanicus, fed the flagellate Isochrysis galbana with varying lipid composition and the diatom Chaetoceros muelleri

The effect of feeding the flagellate Isochrysis galbana (Parke; clone T-Iso) of modified lipid composition on the growth and lipid composition of juvenile scallops [Placopecten magellanicus (Gmelin)] was investigated in the spring of 1993. I. galbana grown in 85-liter cage culture turbidostats under conditions of nitrogen limitation had a significantly higher total lipid content than when grown under nutrient-replete conditions. This was due mainly to a doubling in the amount of less unsaturated triacylglycerol in the cells. The concentrations of methyl and ethyl ketones were also greater in nitrogen-limited cells. Diets of nitrogen-limited I. galbana and nutrient-replete I. galbana grown in continuous and semi-continuous cultures were compared. Scallop juveniles were batch fed daily, and measurements of ingestion were determined. Samples of juveniles were removed periodically for determination of organic weight. The juveniles did not grow when fed nitrogen-limited or nutrient-replete I. galbana alone; however, when each diet was supplemented with 20% of the diatom Chaetoceros muelleri (Lemm.), there was a significant increase in growth in the juveniles receiving the nitrogen-limited I. galbana compared with juveniles on other diets. In comparison with I. galbana,  C. muelleri provided a rich source of carbohydrates and the essential fatty acid 20:4ω6. This study shows the importance of providing optimal dietary levels of ω3 and ω6 polyunsaturated fatty acids, as well as less unsaturated fatty acids and carbohydrates. Received: 29 September 1997 / Accepted: 2 October 1998     

Ontogenic variations in fatty acid and alcohol composition of the pelagic amphipod <Emphasis Type="Italic">Themisto libellula</Emphasis> in Kongsfjorden (Svalbard)

The fatty acid and alcohol composition of the pelagic amphipod, Themisto libellula, was monitored during the 5 first months of its life cycle (4–20 mm length) in an Arctic fjord, Kongsfjorden, Svalbard. Fatty acids of the three major lipid classes, polar lipids (PL), triacylglycerol (TAG), and wax esters (WE), were analyzed to highlight ontogenic changes in their diet and metabolism. The PL composition of T. libellula did not show any strong variations along their growth except during the first month where an important increase of 20:5(n-3) (EPA) and 22:6(n-3) (DHA) was observed. The TAG composition revealed a clear gradient corresponding to a diet shift from omnivorous juveniles toward carnivorous sub-adults and adults. Indeed, fatty acid trophic markers of diatoms were dominant in the juveniles, whereas 20:1(n-9) and 22:1(n-11), the Calanus sp. trophic markers, overwhelmed in the older stages. The WE composition highlighted the same general trend, however, differences were found with the TAG and are discussed as a result of differences in turnover rates and assimilation pathways between the two lipid classes.     

Using fatty acid analysis to elucidate the feeding habits of Southern Ocean mesopelagic fish

Fatty acid biomarkers were used to investigate the feeding ecology of 17 mesopelagic fish species occurring in the Southern Ocean. Fatty acid signatures of species where little or no dietary information exists were compared to fatty acid signatures of species of known diets in order to elucidate their trophic position. Principal component analysis grouped species of known diets into two clusters with amphipods and copepods comprising the main prey species, respectively. Although the majority of species of unknown diet were grouped with either of these feeding guilds, a third cluster comprising only Gymnoscopelus bolini was identified suggesting a significantly different diet for this species. Electrona antarctica also exhibited significant changes in fatty acid signatures with size. Furthermore, discriminant analysis of the four most abundant species classified species with a 90% success rate thus validating the usefulness of fatty acid signatures when trying to resolve the trophic position of species where no or little dietary information exists.     

The ecological significance of lipid/fatty acid synthesis in developing eggs and newly hatched larvae of Pacific cod (Gadus macrocephalus)

The lipid/fatty acid composition of marine fish eggs and larvae is linked with buoyancy regulation, but our understanding of such processes is largely restricted to species with pelagic eggs. In this study, we examined developmental changes in the lipid/fatty acids of eggs and embryos of Pacific cod (Gadus macrocephalus), a species that spawns demersal eggs along coastal shelf edges, but as larvae must make a rapid transition to the upper reaches of the water column. Adult Pacific cod were collected in the Gulf of Alaska during the spawning season and eggs of two females were artificially fertilized with sperm from three males for each female. The eggs were subsequently reared in the laboratory to determine (1) how lipids/fatty acids were catabolized during egg and larval development, and (2) whether lipid/fatty acid catabolism had measurable effects on egg/embryo density. Eggs incubated at 4°C began hatching after 3-weeks and continued to hatch over a 10-day period, during which there was a distinct shift in lipid classes (phospholipids (PL), triacyglycerols (TAG), and sterols (ST)) and essential fatty acids (EFAs: 22:6n-3 (DHA), 20:5n-3 (EPA), and 20:4n-6 (AA)). In the egg stage, total lipid content steadily decreased during the first 60% of development, but just prior to hatch we observed an unexpected 2–3-fold lipid increase (~6–9 μg individual⁻¹) and a significant drop in egg density. The increase in lipids was largely driven by PL, with evidence of long-chained fatty acid synthesis. Late-hatching larvae had progressively decreasing lipid and fatty acid reserves, suggesting a shift from lipogenesis to lipid catabolism with continued larval development. Egg density measures suggest that lipid/fatty acid composition is linked to buoyancy regulation as larvae shift from a demersal to a pelagic existence following hatch. The biochemical pathway by which Pacific cod are apparently able to synthesize EFAs is unknown, therefore representing a remarkable finding meriting further investigation.     

Studies on the nutrition of marine flatfish. Growth of the plaice Pleuronectes platessa on diets containing proteins derived from plants and other sources

The growth of 0-group Pleuronectes platessa L. on diets containing plant proteins was examined. Diets were made acceptable to the fish by including in them cod flesh which had been predigested with proteolytic enzymes, as well as a proportion of air-dried cod meal. A leaf-protein concentrate was fed at 6 levels ranging from 0 to 63 % of the total dietary protein; B.P. protein concentrate (Toprina) and soyabean meal were examined at a single level — about 45 % of the dietary protein. The remainder of the protein in these diets was animal protein, cod meal or cod flesh. Analysis of the weight gains after a 12 week feeding period and of the growth curves showed that, as the proportion of leaf protein in the diet increased, the growth rate diminished, the effect becoming more pronounced when leaf protein reached 40 % or more of the dietary protein. Growth on the diet containing B.P. protein concentrate was superior to that on the diet containing soyabean meal and was comparable with diets containing low levels of leaf protein. The protein efficiency ratios (PER= g live weight gain/g protein fed) for the different diets showed similar trends to the growth rates. Diets containing 40 % or less of leaf protein and the B.P. protein diet had PERs of 1.4 to 1.5. The soyabean meal diet and diets containing more than 40 % of the protein as leaf protein had PERs of 1.0 or less. The results are considered in the context of production diets for flatfish.     

Lipid composition of yellowtail flounder (<Emphasis Type="Italic">Limanda ferruginea</Emphasis>) in relation to dietary lipid intake

Condition indices (CI), hepatosomatic indices (HSI) and proximate, lipid and fatty acid composition of wild and laboratory-reared yellowtail flounder (Limanda ferruginea) (Storer, 1839) were compared from data taken throughout spring and summer 1996–1998. Cluster analysis was performed on the fatty acid signatures of these two groups along with a commercial diet and several invertebrate species to determine possible feeding patterns in Conception Bay, Newfoundland. HSI and levels of storage fat were significantly higher in the muscle and liver of laboratory-reared yellowtail flounder, indicating an efficient absorption of dietary lipid and an increase in fat deposition. Fatty acid analysis of the liver showed that wild fish contained significantly higher proportions of the essential fatty acids 20:46, 20:53 and 22:63; whereas proportions of 18:1 and 18:26 were significantly higher in all fractions of lipid examined from laboratory-reared fish than they were from wild fish. Polar fractions of lipid were more similar than the neutral fraction of lipid when comparing wild and laboratory-reared fish. Taken together, the differences in CI, HSI, proximate and fatty acid composition suggest that feeding commercial diets to L. ferruginea can cause changes in patterns of lipid deposition and metabolism. Cluster analysis of marine plankton, sedimenting particulate matter, wild invertebrates, the commercial diet and fish tissues showed that the fatty acid signatures of both wild and laboratory-reared yellowtail flounder closely resembled their respective food items. Fatty acid signatures from wild fish were more closely related to plankton and settling particulate matter, suggesting relatively few steps in the food web leading to yellowtail flounder. In addition to the resemblance between fatty acids in the commercial diet and the tissues of laboratory-reared yellowtail flounder, these fish had similar fatty acid signatures to those of wild invertebrates.Communicated by J.P. Grassle, New Brunswick     

Fatty acid evidence for the importance of myctophid fishes in the diet of king penguins, Aptenodytes patagonicus

We tested the usefulness of the fatty acid signature-method in investigating the diet of seabirds in conjunction with the conventional technique of stomach-content analysis. We compared the fatty acid composition of subcutaneous white adipose tissue (SWAT) of king penguin chicks (Aptenodytes patagonicus) during fattening periods to that of total lipids from their food. In both spring and autumn, the fatty acid composition of chick SWAT was identical to that of the dietary lipids. Because the diet of adult king penguins feeding for self-maintenance (i.e. not for their chicks) was essentially unknown, we subsequently analysed their SWAT fatty acid patterns after premolting and prebreeding foraging trips (during which they build up large energy reserves). The fatty acid composition of SWAT from adults was identical to that of chick adipose tissue and food. King penguin diet and SWAT were characterized by high levels of very long-chain mono-unsaturated fatty acids (20 to 24 carbon atoms, 16 to 23% by mass) and (n-3) poly-unsaturated fatty acids (19 to 27%); these consisted mainly of 20:1n-9 (5 to 8%) and 22:1n-11 (5 to 8%), and 22:6n-3 (10 to 13%) and 20:5n-3 (3 to 9%), respectively. Prey items identified from chick stomach contents indicated that the bulk of the food was oceanic myctophid fishes, mainly Electrona carlbergi, Krefftichthys anderssoni and Protomyctophum tenisoni. The fatty acid composition of four other species of myctophid fishes was similar to that of penguin diet and SWAT, but markedly different from that measured for a squid species and that reported for crustaceans. These findings indicate that adult king penguins prey on myctophid fish not only to feed their chicks but also for their own nutrition. The fatty acid signature-technique is therefore a reliable method to gain information on the food and feeding ecology of seabirds when more conventional techniques are of limited value. Such information is important to the understanding of trophic relationships between key species of the ecosystems, and also to provide insight into the nature of avian adaptations to the marine environment. Received: 11 December 1997 / Accepted: 25 July 1998     

Assimilation of the wax esters of marine zooplankton by herring (Clupea harengus) and rainbow trout (Salmo gairdnerii)

Herring (Clupea harengus L.) and rainbow trout (Salmo gairdnerii) fed frozen-thawed zooplankton for 7 days assimilated more than 90% of the dry matter in the zooplankton consumed. Less than 5% of the total lipid ingested, largely wax esters, was excreted in the faeces. Faecal lipids from both the herring and the trout, especially the latter, were rich in wax esters and free fatty alcohols. Faecal fatty alcohols were enriched in 22:1 and deficient in 16:0 and 16:1 alcohols compared to zooplankton fatty alcohols. The growth of rainbow trout fed frozen-thawed zooplankton over 3 months was about one-half of that of trout fed commercial pellets. The fish fed on zooplankton appeared normal and were more pigmented than the fish fed on commercial pellets.